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Multifactorial role of HIV-vpr in cell apoptosis revealed by a naturally truncated 54aa variant

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Chinese Medical Journal 2021年第7期134卷 845-847页

作者： Ling Du Cui-Song Wu Jun Sun Tong Yu Pan-Pan Lyu San-Feng Han Chao Qiu Zhe-Feng Meng Key Laboratory of digestive cancer full cycle monitoring and precise intervention of Shanghai Municipal Health Commission Minhang HospitalFudan UniversityShanghai 201199China Department of infection the Third People’s Hospital of Zhenjiang CityZhenjiangJiangsu 212021China Institutes of Biomedical Sciences Shanghai Medical CollegeFudan UniversityShanghai 200032China.

Although antiretroviral therapy(ART)is effective at suppressing the human immunodeficiency virus type 1(HIV-1)replication,HIV-1 infection is still a global public health problem.HIV-1 accessory protein viral protein R(vpr)is a multifunctional protein with a primary role in regulating cellular apoptosis.[1]The amino acid(aa)positions 52-96 in the C-terminus of vpr were identified as the apoptosis-regulating domain.[2,3]In this study,we found a natural vpr variant truncated at the 54 aa position(*54vpr)from HIV patients in the acquired immune deficiency syndrome(AIDS)phase that mediated both pro-and antiapoptotic cellular effects by interacting with distinct adenine nucleotide translocator(ANT)isoforms.A novel apoptosis-regulating domain was further identified in the 23-37 aa position in the N-terminus of vpr.

关键词： vpr apoptosis HIV

Programmable activation of Bombyx gene expression using CRISPR/dCas9 fusion systems

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Insect Science 2019年第6期26卷 983-990页

作者： Xiao-Gang Wang San-Yuan Ma Jia-Song Chang Run Shi Ruo-Lin Wang Ping Zhao Qing-You Xia State Key Laboratory of Silkworm Genome Biology Southwest University ChongqingChina Chongqing Engineering and Technology Research Center for Novel Silk Materials Southwest University Chongqing China

The recently developed clustered regularly interspaced short palindromic repeats (CRISPR)-based techniques have made it possible to reprogram target gene expression without cloning complementary DNA or disturbing genomic sequence in mammalian cells and several multicellular organisms. We previously showed that CRISPR-associated protein 9 (Cas9) and CRISPR from Prevotella and Francisella 1 (Cpfl) could induce target mutations, deletions, inversions, and duplications both singly and multiplex in silkworm, Bombyx mori. However, it remains unknown whether the CRISPR activation (CRISPRa) system can be used in B. mori. In this study, we investigated the CRISPRa system, in which a nuclease dead Streptococcus pyogenes Cas9 (SpCas9) is fused to two transcription activation domains, including VP64 (a tetramer of the herpes simplex VP 16 transcriptional activator domain), and vpr (a tripartite activator, composed of VP64, p65, and Rta). The results showed that both dCas9-VP64 and dCas9-vpr systems could be used in B. mori cells, of which the latter showed significantly higher activity. The dCas9-vpr system showed considerable activity on all five tested target genes, and further analysis revealed that the up-regulation of genes was negatively correlated to their basal expression level. We also observed that this system could be used to upregulate a range of target genes. Taken together, our findings demonstrate that CRISPRa can be a powerful tool to study gene functions in B. mori and perhaps other non-drosophila insects.

关键词： Bombyx mori CRISPR/dCas9 system transcriptional activation VP64 vpr

Viral infections and cell cycle G2/M regulation

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Cell Research 2005年第3期15卷 143-149页

作者： Richard Y. ZHAO,Robert T. ELDER Children's Memorial Research Center Northwestern University Feinberg School of Medicine Chicago IL 60614 USA

Progression of cells from G2 phase of the cell cycle to mitosis is a tightly regulated cellular process that requires activation of the Cdc2 kinase, which determines onset of mitosis in all eukaryotic cells. In both human and fission yeast (Schizosaccharomyces pombe) cells, the activity of Cdc2 is regulated in part by the phosphorylation status of tyrosine 15 (Tyr15) on Cdc2, which is phosphorylated by Wee1 kinase during late G2 and is rapidly dephosphorylated by the Cdc25 tyrosine phosphatase to trigger entry into mitosis. These Cdc2 regulators are the downstream targets of two well- characterized G2/M checkpoint pathways which prevent cells from entering mitosis when cellular DNA is damaged or when DNA replication is inhibited. Increasing evidence suggests that Cdc2 is also commonly targeted by viral proteins, which modulate host cell cycle machinery to benefit viral survival or replication. In this review, we describe the effect of viral protein R (vpr) encoded by human immunodeficiency virus type 1 (HIV-1) on cell cycle G2/M regulation. Based on our current knowledge about this viral effect, we hypothesize that vpr induces cell cycle G2 arrest through a mechanism that is to some extent different from the classic G2/M checkpoints. One the unique features distinguishing vpr-induced G2 arrest from the classic checkpoints is the role of phosphatase 2A (PP2A) in vpr-induced G2 arrest. Interestingly, PP2A is targeted by a number of other viral proteins including SV40 small T antigen, polyomavirus T antigen, HTLV Tax and adenovirus E4orf4. Thus an in-depth understanding of the molecular mechanisms underlying vpr-induced G2 arrest will provide additional insights into the basic biology of cell cycle G2/M regulation and into the biological significance of this effect during host-pathogen interactions.

关键词： HIV-1 vpr cell cycle G2/M regulation DNA damage DNA replication checkpoints viral infections fission yeast Schizosaccharomyces pombe.

病毒蛋白调控逆转录元件Ⅰ型长弥散核元件活性的影响及其意义

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中国生物制品学杂志 2023年第1期36卷 98-104页

作者：赵梓含王宇(综述) 赵一霏赵可(审校) 吉林大学白求恩第一医院转化医学研究院艾滋病与病毒研究所吉林长春130061

型长弥散核元件(long interspersed element-1,LINE-1)是已知人类细胞中唯一具有活性的、可自主转座的逆转录元件(retroelements),与自身免疫性疾病相关,对细胞的抗病毒天然免疫,特别是对细胞干扰素(interferon,IFN)水平起到重要的激活... 详细信息

型长弥散核元件(long interspersed element-1,LINE-1)是已知人类细胞中唯一具有活性的、可自主转座的逆转录元件(retroelements),与自身免疫性疾病相关,对细胞的抗病毒天然免疫,特别是对细胞干扰素(interferon,IFN)水平起到重要的激活、调控作用。本文阐述了包括人类免疫缺陷病毒(human immunodeficiency virus,HIV)、乙型肝炎病毒(hepatitis B virus,HBV)等病毒的几种非结构蛋白参与调控LINE-1活性的机制。这些机制既保证了病毒基因组的正常表达,又参与调控细胞天然免疫水平,对相关机制的抑制可能为治疗病毒感染引起的疾病提供新的方法。

关键词：型长弥散核元件 Nef vpr Vpx HBx

RelB对HIV-1 vpr转录激活及诱导细胞G2/M期停滞功能影响的初步研究

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病毒学报 2013年第1期29卷 44-50页

作者：刘瑞康高杨林永权谈娟耿运琪乔文涛南开大学分子微生物与技术教育部重点实验室南开大学生命科学学院天津300071

vpr是人类免疫缺陷病毒(Human Immunodeficiency Virus type 1,HIV-1)的辅助蛋白之一,在病毒复制及AIDS进程中起重要作用。为了研究vpr完成其生物学功能的分子机制,本研究利用酵母双杂交技术,从人的cDNA文库中筛选,并经免疫共沉淀技术证... 详细信息

vpr是人类免疫缺陷病毒(Human Immunodeficiency Virus type 1,HIV-1)的辅助蛋白之一,在病毒复制及AIDS进程中起重要作用。为了研究vpr完成其生物学功能的分子机制,本研究利用酵母双杂交技术,从人的cDNA文库中筛选,并经免疫共沉淀技术证实NF-κB通路中的重要蛋白RelB与vpr存在相互作用;发现RelB蛋白能促进vpr介导的对NF-κB报告基因的激活,也能促进vpr对HIV-1LTR的反式激活作用。利用流式细胞技术发现RelB促进vpr诱导细胞周期G2/M期停滞。上述结果表明,RelB辅助vpr完成其转录激活以及调控细胞周期的功能。

关键词： vpr RelB 转录激活细胞G2 M停滞

HIV-1 vpr蛋白对C8166细胞毒性研究

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中华微生物学和免疫学杂志 2008年第10期28卷 899-903页

作者：贺芳曾耀英王通肇静娴臧宁林长乐暨南大学组织移植与免疫实验室广州510632

目的利用腺病毒系统研究HIV-1vpr蛋白在T细胞来源的C8166细胞内的高表达以及细胞内该蛋白对T细胞毒性。方法将表达目的蛋白vpr的重组腺病毒rAd—vpr和空白载体病毒rAd—vector分别感染对数生长期的C8166细胞，流式细胞术（FCM）检测细... 详细信息

目的利用腺病毒系统研究HIV-1vpr蛋白在T细胞来源的C8166细胞内的高表达以及细胞内该蛋白对T细胞毒性。方法将表达目的蛋白vpr的重组腺病毒rAd—vpr和空白载体病毒rAd—vector分别感染对数生长期的C8166细胞，流式细胞术（FCM）检测细胞周期分布、凋亡和坏死。用Hoechst—PI荧光染色观察细胞的凋亡和坏死，JC-1荧光染色测定线粒体膜电势。结果An—nexinV—PI染色及Hoechst—PI染色一致显示HIV-1vpr能显著诱导C8166细胞凋亡和坏死；PI细胞周期结果表明HIV-1vpr能阻滞C8166细胞于G2期；JC—1荧光染色法测定HIV-1vpr致C8166线粒体膜电势下降。结论细胞内HIV-1vpr介导的T细胞毒性包括线粒体功能障碍、细胞周期的G2期阻滞和细胞的死亡。

关键词：重组腺病毒 HIV-1 vpr C8166细胞细胞死亡

基于正态分布的一种岩石（岩块）弹性纵波速度取值方法

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物探化探计算技术 2023年

作者：洪明宏张军鹏左国青中国电建集团中南勘测设计研究院有限公司

新鲜完整岩石(岩块)波速（vpr）是计算岩体完整性系数的重要参数，岩石弹性纵波速度取值对岩体完整性评判具有重要作用，但其取值具有较强人为随意性的缺点，对同一类型岩石原位测试进行统计分析，得到了岩石弹性纵波速度分布的概率分... 详细信息

新鲜完整岩石(岩块)波速（vpr）是计算岩体完整性系数的重要参数，岩石弹性纵波速度取值对岩体完整性评判具有重要作用，但其取值具有较强人为随意性的缺点，对同一类型岩石原位测试进行统计分析，得到了岩石弹性纵波速度分布的概率分布模型。通过不同概率保证率，可以利用概率分布模型得到完整岩体波速的上限值，得到岩石弹性纵波速度参数取值范围。以某抽水蓄能电站中花岗岩试验数据为例，通过实际工程算例表明，所提出的基于正态分布的岩体弹性波参数取值的方法可为实际工程中确定岩体完整性评价提供参考。

关键词： vpr 正态分布弹性波波速

Two Highly Variable vpr<sub>84</sub>and vpr<sub>85</sub>Residues within the HIV-1-vpr C-Terminal Protein Transduction Domain Control Transductionnal Activity and Define a Clade Specific Polymorphism

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World Journal of AIDS 2014年第2期4卷 148-155页

作者： Jean-Herve Colle Thiery Rose Christine Rouzioux Alphonse Garcia EA 3620 Universite Paris Descartes AP-HP Laboratoire de Virologie Hopital Necker Paris France Laboratoire E3 Phosphatases Unite Signalisation Moleculaire et Activation Cellulaire Institut Pasteur Paris France Unite d’Immunogenetique Cellulaire Institut Pasteur Paris France Unite de Biologie des Populations Lymphocytaires Institut Pasteur Paris France

The virally encoded HIV-1 viral protein R (vpr) is a multifunctional factor that is required for induced HIV-1 pathogenesis. vpr is also a cell-penetrating protein found in biological fluids from HIV-1 infected individuals. In this regard, we previously published that the C-terminal vpr77-92 sequence from HIV-1 89.6, but not from pNL4.3 strain, is a new pro-apoptotic and protein transduction domain (PTD). Here we report on a sequence analysis of vpr77-92 domain using the Los Alamos HIV-1 sequence database. The analysis showed that the two residues of the domain vpr84 and vpr85 are highly variable and differently biased in HIV-1 clade B and HIV-1 clade C. Furthermore, when Jurkat lymphoblastoid cells or PBMC were incubated with chemically synthesized peptides containing distinct vpr77-92 C-terminal sequences from clades B or C, we found that a clade-dependent polymorphism in vpr84 and vpr85 residues controlled the transducing activity of the C-terminal HIV-1 vpr77-92 domain. Together our data indicate that clade-dependent polymorphism in the vpr84 and vpr85?residues defines the transducing properties mediated by the C-terminal domain of HIV-1 vpr. Identification of this vpr polymorphism suggests new approaches to understand the HIV-1 biology and/or pathogenesis.

关键词： vpr HIV-1 Subtypes Polymorphism Transduction

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创新创业教育评价的vpr结构模型

教育研究 2019年第7期40卷 83-90页

作者：徐小洲浙江传媒学院浙江大学教育学院

评价要素及其结构的确定是创新创业教育评价的重要基础。研究采用扎根理论方法,通过对访谈资料的内容分析,提出vpr三维三级创新创业教育评价的理论结构模型。vpr评价体系由3个一级维度、8个二级维度和29个三级评价维度构成。3个一级维... 详细信息

评价要素及其结构的确定是创新创业教育评价的重要基础。研究采用扎根理论方法,通过对访谈资料的内容分析,提出vpr三维三级创新创业教育评价的理论结构模型。vpr评价体系由3个一级维度、8个二级维度和29个三级评价维度构成。3个一级维度是价值评价(Value)、过程评价(Process)和结果评价(Result);8个二级维度是精神价值、现实价值、政策投入、教育投入、学生发展、企业发展、创业者绩效、产品与成果。该评价模型强调精神价值评价与现实价值评价相结合、发展性评价与绩效性评价相结合、短期评价与长期评价相结合,具有多维、多层、多元的特征,丰富了创新创业教育评价的内容,对创新创业教育实践具有理论参考价值。

关键词：创新创业教育 vpr 评价结构多层次模型